白菜种传黑斑病菌rDNA ITS区序列分析

本文以来自国内外的20株白菜黑斑病菌及近源种为研究材料,进行了5.8SrDNA及其侧翼ITS区的克隆、测序、序列变异及遗传进化关系分析。黑斑病菌及其近源种真菌核糖体5.8SrDNA及其侧翼ITS区序列比对结果显示,不同种菌株ITS1比ITS2在碱基构成上有更大变异,而且ITS1的序列长度变异比ITS2的大;而种内虽然各菌株的寄主和地理来源不同,但ITS1和ITS2在长度上均没有变异,碱基构成上存在微小的变异。对该区序列的聚类分析表明,白菜黑斑病菌3个种芸薹链格孢Alternariabrassicae、甘蓝链格孢A.brassicicola和萝卜链格孢A.japonica虽然地理来源和寄主不同,但种内的不同菌株均在一个独立的聚类组中,种之间以及其和链格孢属内其它种在聚类关系上能明显分开,可基于该区进行黑斑病菌的分类鉴定。     

Alternaria jesenskae sp. nov., a new species from Slovakia on Fumana procumbens (Cistaceae)

Alternaria jesenskae sp. nov. recovered from seeds of a shrubby perennial plant Fumana procumbens (Cistaceae) in Slovakia is described and illustrated. The new taxon can be clearly separated from the other related large-spored and filament-beaked Alternaria species based on sequences of the ITS1, 5.8S and ITS2 region as well as by its distinctive morphology. Even though the molecular data have shown close relatedness with A. multirostrata, the new species is morphologically most similar to A. tomatophila distinguished primarily by the pronounced colony pigmentation, conidial septation and beak branching.     

小麦种子内携真菌多样性研究

为揭示小麦种子所携带真菌的种类和多样性,丰富小麦相关微生物的可利用资源,我们对来自俄罗斯、澳大利亚、哈萨克斯坦、加拿大以及中国的57份小麦种子样品进行内携真菌分离并对其中的21份种子利用高通量测序进行多样性分析.结果显示,分离获得564株真菌,经ITS序列分析结合形态观察鉴定为31属69种,其中菌株数量相对多度最高的5...     

应用5.8S rDNA及ITS区序列分析链格孢种级分类

对9个链格孢小孢子种和3个大孢子种共20个链格孢菌株的5.8S rDNA及其两侧的ITS1区和ITS2区进行了序列分析。聚类分析结果表明形态差异大的种可以明确加以区分,而供试的9个小孢子种之间差异很小,不能根据对所选区段的序列分析加以区分。传统分类上的滨菊链格孢不属于Alternaria, 其分类地位需进一步研究。     

应用5.8S rDNA及ITS区序列分析链格孢种级分类*

对9个链格孢小孢子种和3个大孢子种共20个链格孢菌株的5.8S rDNA及其两侧的ITS1区和ITS2区进行了序列分析。聚类分析结果表明形态差异大的种可以明确加以区分,而供试的9个小孢子种之间差异很小,不能根据对所选区段的序列分析加以区分。传统分类上的滨菊链格孢不属于Alternaria, 其分类地位需进一步研究。     

Emendation of Dioszegia with redescription of Dioszegia hungarica and two new combinations,Dioszegia aurantiaca and Dioszegia crocea

During phylogenetic analyses of hymenomycetous yeasts based on 18S rDNA sequences, we found that Bullera armeniaca showed an extremely close phylogenetic relationship to Cryptococcus hungaricus. The analyses of internal transcribed spacer (ITS) regions of the two yeasts and the phylogenetically related species, Bullera aurantiaca and Bullera crocea, showed that B. armeniaca and C. hungaricus had identical sequences, indicating that these were conspecific. B. aurantiaca and B. crocea also showed high sequence similarity, 97.1% for ITS1, 100% for ITS2, and 98.7% for overall ITS regions. A DNA-DNA reassociation experiment revealed that B. armeniaca and C. hungaricus were conspecific and B. aurantiaca and B. crocea were two distinct species. These species occurred at a phylogenetically different lineage from that of Bulleromyces albus (anamorph: Bullera alba, type species of Bullera) and Filobasidiella neoformans (anamorph: Cryptococcus neoformans, neotype species of Cryptococcus). Based on these results, we emend the genus Dioszegia to include both ballistoconidium-forming and non-ballistoconidium-forming yeasts and redescribe the species Dioszegia hungarica. B. aurantiaca and B. crocea are also transferred to Dioszegia as Dioszegia aurantiaca comb. nov. and Dioszegia crocea comb. nov.     

Alternaria alternata prevalence in cereal grains and soybean seeds from Entre Ríos, Argentina

A mycological survey was carried out at Entre Ríos province, Argentina, on sorghum grain, maize, rice, soybean seeds and on freshly harvested and stored wheat. The isolation frequencies and relative densities of species belonging to genera Alternaria, Aspergillus, Fusarium, Penicillium and other fungi were calculated. Alternaria alternata was the major fungal species isolated from sorghum, rice, soybean seeds and on freshly harvested wheat, and a low incidence of Fusarium species was observed on the same substrates. In maize the major fungal species isolated was Fusarium verticillioides. The high incidence levels of A. alternata observed,suggest that it may be necessary to determine, among other mycotoxins, if Alternaria toxins occur in these commodities.     

一株产色素真菌的形态及分子鉴定

从云南高原地区的竹叶病理组织上分离到1株可产生红棕色素的丝状真菌ZL07-8。通过对分生孢子梗、分生孢子、喙等的形态学研究及对ITS和5.8S rRNA序列的分子系统学分析,发现此菌为链格孢属小孢子种群成员。然而该菌具有区别于已知种的特征,如分生孢子梗常单生,少数成簇,无分枝;分生孢子大多单生,较少链生,孢身较大;寄主为华西箭竹,故立为竹链格孢新种。干制培养物ZL07-8D保藏于山东理工大学生命科学学院实验室。     

蒟蒻薯属 (薯蓣科) 植物DNA条形码研究

蒟蒻薯属（Tacca）植物种间在形态上差别不大,导致分类上存在一定的困难。DNA条形码是一种利用短的DNA标准片段来鉴别和发现物种的方法。本研究利用核基因ITS片段和叶绿体基因trnH psbA, rbcL, matK片段对蒟蒻薯属6个种的DNA条形码进行研究,对4个DNA片段可用性,种内种间变异,barcode gap进行了分析,采用Tree based和BBA两种方法比较不同序列的鉴定能力。结果显示：单片段ITS正确鉴定率最高,片段组合rbcL+matK正确鉴定率最高。支持CBOL植物工作组推荐的条码组合rbcL+matK可作为蒟蒻薯属物种鉴定的标准条码,建议ITS片段作为候选条码。丝须蒟蒻薯Tacca integrifolia采自西藏的居群与马来西亚居群形成了2个不同的遗传分支,且两者在形态上也存在一定的差异,很可能是一个新种。     

Horizontal gene and chromosome transfer in plant pathogenic fungi affecting host range

Plant pathogenic fungi adapt quickly to changing environments including overcoming plant disease resistance genes. This is usually achieved by mutations in single effector genes of the pathogens, enabling them to avoid recognition by the host plant. In addition, horizontal gene transfer (HGT) and horizontal chromosome transfer (HCT) provide a means for pathogens to broaden their host range. Recently, several reports have appeared in the literature on HGT, HCT and hybridization between plant pathogenic fungi that affect their host range, including species of Stagonospora/Pyrenophora, Fusarium and Alternaria. Evidence is given that HGT of the ToxA gene from Stagonospora nodorum to Pyrenophora tritici-repentis enabled the latter fungus to cause a serious disease in wheat. A nonpathogenic Fusarium species can become pathogenic on tomato by HCT of a pathogenicity chromosome from Fusarium oxysporum f.sp lycopersici, a well-known pathogen of tomato. Similarly, Alternaria species can broaden their host range by HCT of a single chromosome carrying a cluster of genes encoding host-specific toxins that enabled them to become pathogenic on new hosts such as apple, Japanese pear, strawberry and tomato, respectively. The mechanisms HGT and HCT and their impact on potential emergence of fungal plant pathogens adapted to new host plants will be discussed.     

The taxonomy of the Pollenia rudis species-group in the Holarctic Region (Diptera: Calliphoridae)

Abstract. A rudis species-group is defined within Pollenia Robineau-Desvoidy, and new characters found useful in the taxonomy of this genus are presented. P.rudis (Fabricius), P.angustigena Wainwright, stat.rev. and P.pseudorudis Rognes are redescribed. P.hungarica sp.n., P.longitheca sp.n. and P.luteovillosa sp.n. are described as new to science. A key is provided, and the terminalia of both sexes are illustrated for all the species. Some features of the puparia are figured for the species where these are known. A neotype is designated for Musca rudis , and a lectotype for P.angustigena. P.angustigena, P.pseudorudis and P.rudis are Holarctic species, and the latter two have also been found in New Zealand. The remaining species are confined to the western Palaearctic. P.hungarica is known from central Europe, including southern parts of Scandinavia, P.longitheca from the eastern Mediterranean, and P.luteovillosa from Algeria and Morocco in North Africa. In the larval stages P. rudis group members are parasites of or predators on earthworms. The species have several generations each year, and normally overwinter as adults. Eisenia rosea (Savigny) serves as a host for P.hungarica, P.pseudorudis and P.rudis according to the reared material available. A previous detailed account of the immature stages and life-cycle of lsquo;rudis' from North America is tentatively assigned to pseudorudis. Keilin's (1909,1915) often cited accounts of the immature stages and life-cycle of a species called 'rudis' are rejected as a source of information for any member of the rudis group.     

北京地区非洲菊叶斑病病原菌鉴定

采用离体叶片接种致病性测定、形态学观察和分子鉴定方法对非洲菊叶斑病病原进行鉴定。从北京某花卉基地采集非洲菊叶斑病样,分离获得22个真菌分离物,分别记作GL1-GL22。经过致病性试验证实,分离物GL14和GL22为非洲菊叶斑病病原菌。经形态学鉴定,GL14为细极链格孢Alternaria tenuissima,GL22为链格孢Alternaria alternata。采用真菌通用引物对GL14和GL22 rDNA的ITS区进行PCR扩增,并将测序结果在GenBank中进行同源性比对分析,分子鉴定与形态学鉴定结果一致。     

基于rDNAITS序列分析Alternaria与相似属的关系及A.leucanthemi的分类地位

对Alternaria,Stemphylium,Ulocladium,Nimbya,Embellisia等五个形态相似属的代表性种进行了5.8S/ITS区段序列测定,连同从GenBank下载的相关有性型Pleospora,Lewia的同项资料,由Neighbor-joining方法构建系统发育树。序列对比结果显示,所有参试的菌株/种的5.8SrDNA序列保守程度较高,ITS序列变化较大,ITS1比ITS2变化更大。由5.8S/ITS构建的系统发育树可以足够将供试菌区分到属;系统发育树显示Alternaria,Ulocladium,Nimbya和Embellisia系统学关系较近,又存在一定的分化;Stemphylium聚在相对较远的分支上,与上述四属关系较远,相对独立进化;有性型的Pleospora与无性型的Stemphylium被聚到一个分支,这种不同阶段菌株在遗传基础上的一致,充分证明所用方法及所选测序的rDNA区段对本类群真菌的分类很有价值。Alternarialeucanthemi位于系统发育树的外侧,较为独立进化,与Alternaria的其它种亲缘关系较远,却与Stemphyliumspp.和Bipolarisspp.关系较近,对其目前的分类地位提出质疑。     

Characterisation of a new species of Pythium isolated from a wheat field in northern France and its antagonism towards Botrytis cinerea causing the grey mould disease of the grapevine

A new species, Pythium bifurcatum, isolated from soil samples taken from a wheat field in Lille in northern France is described here. The oomycete occurred thrice out of 50 samples. The type specimen is F-91, which is a slow-growing saprophyte living on vegetable debris and which can be recognised by its antheridial as well as oogonial characteristics, which are different from other known species of Pythium. When grown together with Botrytis cinerea, the causal agent of the grey mould disease of the grapevine, Pythium bifurcatum shows a pronounced antagonism and suppresses its growth. Morphological features of this new species, its antagonism to B. cinerea, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed in this article.     

Alternaria capsicicola sp. nov., a new species causing leaf spot of pepper (Capsicum annuum) in Malaysia

A new species of Alternaria causing leaf spot of pepper (Capsicum annuum) obtained from the Cameron highlands, Pahang, Malaysia, was determined based on phylogenetic analyses, morphological characteristics, and pathogenicity assays. Phylogenetic analyses of combined dataset of the glyceraldehyde-3-phosphate dehydrogenase (gpd), Alternaria allergen a 1 (Alt a1) and calmodulin genes revealed that the new isolates clustered into a subclade distinct from the closely related Alternaria species A. tomato and A. burnsii. The solitary or short chains of conidia resemble those of A. burnsii. However, conidia with long beaks are morphologically similar to A. tomato. Hence, the pathogenic fungus is proposed as Alternaria capsicicola sp. nov. Pathogenicity assays indicated that A. capsicicola causes leaf spot on pepper.     

Pythium abappressorium--a new species from eastern Washington

A new species of Pythium isolated from wheat and apple roots in eastern Washington is described. Pythium abappressorium sp. nov. is characterized by abundant appressoria. Plerotic oospores and sporangia are formed from the appressoria and remnants of the appressoria remain attached to the base of sporangia at maturity. Smaller appressorial swellings, reminiscent of hyphal swellings, are also formed within the appressoria. Pythium abappressorium is pathogenic to wheat, causing damping-off and stunting, but is not pathogenic to apples. The fungus can grow in the temperature range 5 to 30 C, with an optimum of 20 C. The sequence of the ITS1 region of the rDNA did not match the sequences from a worldwide collection of over 1200 isolates, including types and neotypes, suggesting that this species has not been previously described.     

Differentiation of Listeria isolates by PCR amplicon profiling and sequence analysis of 16S-23S rRNA internal transcribed spacer loci

The 16S-23S rRNA internal transcribed spacer region (ITS) in genomic DNA from Listeria species was amplified and sequenced so as to find sequence differences that would allow rapid species and strain differentiation. Agarose gel profiles of amplicons generated with primers designed to amplify ITS loci indicated that Listeria DNA can contain at least two distinct ITS regions. The direct sequencing of the smaller of these ITS amplicons (330 bp) was found useful for the rapid and accurate differentiation of various Listeria species. On the other hand analysis of ITS amplicons generated from a total of 27 L. monocytogenes strains indicated that 4/27 of these strains could be distinguished on the basis of their ITS profile (the presence of a unique 350 bp amplicon). The lack of sequence heterogeneity in the small 333 bp amplicon did not permit rapid strain differentiation.     

生于豆科植物上的链格孢属真菌一新种

报道生于豆科植物沙冬青Ammopiptanthus mongolicus上的链格孢属真菌一新种,沙冬青链格孢Alternaria ammopiptanthi。此种不同于已从豆科植物上报道的5个长喙链格孢种（复喙链格孢A. multirostrata、决明链格孢A. cassiae、猪屎豆生链格孢A. crotalariicola、瓜尔豆链格孢A. cyamopsidis和长喙链格孢A. longirostrata）,主要是其分生孢子的长喙不分枝和孢身细瘦。研究过的模式标本（PSNXAAFS 267852）保存在宁夏农林科学院植物病害标本室。     

链格孢属小孢子种的RAPD分析*

用筛选出的12个随机引物,对链格孢属Alternaria13个小孢子种和作为对照的3个大孢子种共55个分离系(isolates)进行RAPD分析。大孢子种Alternariasolani、A.porri和形态独特的A.leucanthemi在树状聚类图上遗传距离0.44处与所有供试小孢子种区分开,它们彼此之间在遗传距离0.25处相区分,表明所采用的RAPD分析方法适于链格孢种间亲缘关系的研究。所有供试的链格孢小孢子种在遗传距离0.31水平上被聚在一起,表明它们之间的亲缘关系较近。A.infectoria与其它链格孢小孢子种之间遗传距离较远;A.longipes的3个分离系和A.brassicicola的7个分离系在较低的遗传距离上被聚在一起,表明它们是独立的种。其它供试链格孢小孢子种的不同分离系在树状聚类图上未显示明确区分。